KR20140132621A - A stabilized pemetrexed preparation - Google Patents

Abstract

The present invention relates to a stabilized pemetrexed preparation, and more specifically, to a stabilized pemetrexed preparation containing acetylcysteine as an antioxidant and sodium citrate as a buffer. Furthermore, the present invention relates to a pemetrexed preparation which is placed within a sealed container and which contains, as an active ingredient, pemetrexed or a pharmaceutically acceptable salt thereof, wherein the oxygen content in the head space within the container is 3 v/v% or less.

Description

Stabilized pemetrexed preparation < RTI ID = 0.0 >

The present invention relates to a stabilized pemetrexed formulation, and more particularly to a stabilized pemetrexed formulation comprising acetylcysteine as an antioxidant and sodium citrate as a buffer. The present invention also relates to a pemetrexed preparation comprising, as an active ingredient, pemetrexed or a pharmaceutically acceptable salt thereof, contained in a sealed container, wherein the amount of Pemetrake having an oxygen content in the headspace of 3 v / v% Lt; / RTI >

Compounds known to have certain antifolate activities are well known as chemotherapeutic agents for the treatment of cancer. U.S. Patent No. 5,344,932 describes a process for the preparation of certain substituted pyrrolo [2,3-d] pyrimidine-based antifloxate derivatives containing femetrexed, in EP 0434426 a series of 4 -Hydroxypyrrolo [2,3-d] pyrimidine-L-glutamic acid derivatives are known.

Pemetrexed is a 5-substituted pyrrolo [2,3-d] pyrimidine that metabolizes folate metabolites in a variety of cancers, including non-small cell lung cancer Is a multitargeted antifolate that exhibits anticancer efficacy by inhibiting the activity of the antifolate.

Pemetrexed was introduced into cells via reduced folate carrier (RFC), which is the main intracellular transport channel of folic acid, and then converted into polyglutamate (FPGS) by folylpolyglutamate synthetase (FPGS) ) Derivatives and is known to target thymidylate synthase (TS) and dihydrofolate reductase (DHFR).

At present, (ALIMTA ) Is commercially available as a therapeutic agent for the treatment of non-small cell lung cancer and malignant pleural mesothelioma it has been developed under the name of that reference ^{(Physicians' Desk Reference, 60 th} ed., Pp. 1722-1728 (2006)). The lyophilized formulation, which should be reconstituted with 0.9% sodium chloride solution upon administration to the patient and finally to be diluted with 0.9% sodium chloride solution (0.25 mg / ml final concentration) 100 or 500 mg).

Formulations in the form of lyophilized powder are complex in their manufacturing process and require high processing costs. In addition, there is a risk of contamination by microorganisms upon reconstitution of the lyophilized preparation, and pharmacists, doctors, nurses, and the like involved in the preparation of the drug are likely to be exposed to the cell destruction materials. Therefore, in the case of cytotoxic anticancer drugs such as pemetrexed, it is necessary to develop a ready-to-use liquid preparation that can be stored for a long period of time rather than a freeze-dried preparation.

In many cases, the problem with liquid formulations is instability during storage. Due to this instability, a large number of injections are used in the form of lyophilized preparations which are used by dissolving immediately before injection. Even in the case of pemetrexed or its pharmacologically acceptable salt, which is started to be provided as a lyophilized formulation, the preparation of an aqueous solution increases the amount of unknown suppositories, which poses a problem of stability that can not be stored at room temperature for a long period of time. Because of this stability problem, it is currently being used as a lyophilized formulation in clinical practice.

Several formulations have been proposed to overcome these disadvantages. For example, U.S. Patent No. 6,686,365 (Korean Patent Publication No. 2002-0081293) discloses a stable solution of a Pemetrexed solution containing an effective therapeutic amount of Pemetrexed, an effective amount of an antioxidant, and a pharmaceutically acceptable excipient. Glycerol, L-cysteine, and thioglycolic acid.

However, as a problem of the above-mentioned patent, it has been reported that stability is not guaranteed for a desired period of time due to problems such as precipitation when the long-term stability of 25 ° C progresses (International Patent No. WO2012 / 015810) There has been no experimental or commercial realization of stable Pemetrexed liquid preparations for long-term storage. Actually, the inventors of the present invention conducted a stability test after preparation of a liquid preparation containing pemetrexed using L-cysteine, which is the antioxidant mentioned above. As a result, And lowering of the pH. The present inventors also prepared liquid formulations containing pemetrexed using about 60 kinds of stabilizers such as ascorbic acid, sodium thiosulfate, butylated hydroxyanisole, propyl gallate, EDTA, L-methionine and acetylcysteine All of these were unsuitable for stability.

Under these circumstances, the present inventors have conducted extensive studies to overcome the stability problem of the pemetrexed liquid formulation. As a result, it has been found that when acetyl cysteine as an antioxidant and sodium citrate as a buffer are used together, the phentetralis isomer and The present inventors have completed the present invention by confirming that it is possible to effectively reduce or eliminate the occurrence of unknown suppositories and to produce a pemetrexed preparation in which a high stability is maintained during a storage period in a state of a clear solution in which no precipitate is formed . Further, the inventors of the present invention have confirmed that the stability of the preparation can be ensured by controlling the amount of oxygen in the head space, that is, the space above the container when mass-producing the preparation.

It is an object of the present invention to provide a stabilized pemetrexed formulation.

In order to solve the above problems, the present invention provides a pemetrexed preparation comprising Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate as an active ingredient.

In the present invention, the concentration ratio of the above-mentioned Pemetrex or its pharmaceutically acceptable salt, N-acetyl-L-cysteine and sodium citrate may be 1 to 30: 0.15 to 2.0: 1.0 to 15.0.

Preferably, the concentration ratio of the Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate may be 1 to 30: 1.5: 1.0 to 15.0.

In the present invention, the preparation may be a liquid preparation which can be stored in a solution state.

In the present invention, the formulation may be a injectable liquid formulation in a ready-to-use sealed container.

However, the constitution of the present invention is not necessarily limited to injectable liquid preparations, but includes other liquid preparations and non-liquid preparations which can be constituted according to the methods commonly used in the art or those skilled in the art.

The present invention also relates to a pemetrexed formulation comprising, as an active ingredient, pemetrexed or a pharmaceutically acceptable salt thereof, contained in a sealed container, said formulation having an oxygen content in the headspace within the container of 3 v / v% (v / v = oxygen volume / headspace volume, the same applies below).

In the present invention, it is preferable that the preparation further comprises N-acetyl-L-cysteine and sodium citrate.

In the present invention, the concentration ratio of the above-mentioned Pemetrex or its pharmaceutically acceptable salt, N-acetyl-L-cysteine and sodium citrate may be 1 to 30: 0.15 to 2.0: 1.0 to 15.0.

Preferably, the concentration ratio of the Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate may be 1 to 30: 1.5: 1.0 to 15.0.

In the present invention, the preparation may be a liquid preparation which can be stored in a solution state.

In the present invention, the formulation may be a injectable liquid formulation in a ready-to-use sealed container.

In the present invention, the preparation may be such that oxygen in the headspace in the vessel is replaced with an inert gas to adjust the amount of oxygen to 3 v / v% or less.

In the present invention, the inert gas may be nitrogen or argon, but is not limited thereto.

Hereinafter, the present invention will be described in detail.

In general, there are a wide variety of antioxidants used for the purpose of stabilizing the preparation, and specific examples thereof include parahydroxybenzoic acid ester derivatives, alcohols, phenol derivatives, sodium thiosulfate, thimerosal, acetic anhydride, sodium carboxylate, lauryl sulfate, antioxidants, sulfide compounds, sulfite, cystine, Organic acids such as cysteine, cysteamine, amino acids and ascorbic acid, retinol, tocopherol, butylated hydroxyanisole, (butylated hydroxyanisole).

Korean Patent No. 10-0774366 discloses N-acetylamino acid as an antioxidant of paclitaxel. Korean Patent Laid-Open No. 10-2007-0028331 discloses that monothioglycerol, ethylenediaminetetraacetic acid is an antioxidant of diclofenac composition Lt; / RTI >

In the present invention, the antioxidants were subjected to the stabilization test of the injectable formulation of pemetrexed formulation. As a result, it has been confirmed that the general antioxidants as mentioned above have a limitation in daily use when used as an excipient (antioxidant) which is not a main component in the injection form, and that the stability is not improved by the antioxidant alone. In addition, when ascorbic acid, lactic acid, and the like, which are commonly used in common injections, were used, it was confirmed that these injection solutions showed acid instability such as partial discoloration and / or precipitation (Experimental Example 1 and Experimental Example 2).

Surprisingly, however, the inventors of the present invention have found that when acetylcysteine and sodium citrate are used together in a composition comprising pemetrexed or a pharmaceutically acceptable salt thereof as an active ingredient, , The effective ingredient was not substantially changed and discoloration or precipitate was not generated, and it was found that the stability was appropriate for the criteria (Experimental Example 1 and Experimental Example 2). Acetylcysteine and sodium citrate are all commonly used, and their low cost is also advantageous in terms of commerciality.

The pemetrexed formulations of the present invention comprise pemetrexed or a pharmaceutically acceptable salt thereof as an active ingredient. The pemetrexed according to the present invention includes a pharmacologically effective, or a pharmacologically effective drug by an in-body chemical or enzymatic method. Specifically, the pemetrexed drug itself, a pharmaceutically acceptable salt thereof, have.

As used herein, the term "pemetrexed" is a compound having the name of 5-substituted pyrrolo [2,3-d] pyrimidines, specifically represented by the following formula (1) Refers to multiple target antifolates that exhibit anticancer efficacy in various types of cancer, including malignant pleural mesothelioma.

[Chemical Formula 1]

The above-mentioned pemetrexed inhibits the activity of metabolites involved in folate metabolism in various cancer types including non-small cell lung cancer, malignant pleural mesothelioma, and exhibits anti-cancer efficacy.

As used herein, the term "pharmaceutically acceptable salt" refers to salts prepared according to methods conventional in the art, and such methods of preparation are known to those skilled in the art. In particular, the pharmaceutically acceptable salts include, but are not limited to, salts derived from inorganic and organic acids and bases, which are pharmacologically or physiologically acceptable. Examples of suitable acids include hydrochloric acid, hydrobromic acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, Benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include, but are not limited to, alkali metals such as sodium or potassium, alkaline earth metals such as magnesium.

The terms used in the present invention, "acetylcysteine" is having a name of N- acetyl -L- cysteine (N-Acetyl-L-Cysteine , NAC, C 5 H 9 NO 3 S, CAS number 616-91-1) Means a compound used as an antioxidant in a pemetrexed preparation of the present invention. In the present invention, "acetylcysteine" and "N-acetyl-L-cysteine"

Acetylcysteine as an antioxidant is described in the United States Pharmacopeia (US Pharmacopeia 35 - National Formulary 10, p2069). Acetylcysteine is a precursor of L-cysteine, an amino acid contained in foods. Cysteine is oxidized and easily denatured as an insoluble substance, while acetylcysteine is stable and can not be easily denatured, and is a substance that can be administered orally or intravenously.

As used herein, the term "sodium citrate" refers to the sodium salt of citric acid, which is used as a buffering agent in the pemetrexed preparations of the present invention.

The present invention can provide a Pemetrexed preparation capable of stably storing Pemetrexed at a temperature of 1 占 폚 to 30 占 폚 for a long period of time by using acetyl cysteine as an antioxidant and sodium citrate as a buffer .

In the present invention, the pemetrexed preparation is preferably used when the concentration ratio of pemetrexed or a pharmaceutically acceptable salt thereof: N-acetyl-L-cysteine: sodium citrate is 1 to 30: 0.15 to 2.0: 1.0 to 15.0 And particularly preferably in the case where the concentration ratio is 1 to 30: 1.5: 1.0 to 15.0. At this time, the unit of each concentration may be mg / mL. It was found that the production of the flexible material increased beyond the acceptable standard as the storage time elapsed from the pemetrexed formulation different from the concentration ratio (Experimental Example 1).

In the present invention, the Pemetrexed agent may include a pharmaceutically acceptable carrier and a pH adjusting agent.

In the present invention, the pemetrexed preparation is preferably a liquid preparation that can be stored in a solution state, and more preferably, it is a liquid preparation for injectable use contained in a ready-to-use sealed container. have.

In the present invention, when the pemetrexed preparation is a liquid preparation for injection, the pharmaceutically acceptable carrier is water for injection.

In the present invention, the pH of the liquid formulation for pemetrexed juice may preferably be about 6.0 to 8.0, more preferably about 7.2 to 7.8. The pH of the solution can be adjusted by using an acid such as hydrochloric acid or a base such as sodium hydroxide.

The pemetrexed preparation of the present invention may be free of other additives besides acetylcysteine and sodium citrate as described above, but may additionally contain pharmaceutically acceptable excipients. Examples of pharmaceutically acceptable excipients include known additives such as lactose, dextrose, cyclodextrin and derivatives thereof, sucrose, glycerol, sodium carbonate, etc. Examples of suitable excipients include sodium chloride and mannitol.

When the preparation of the present invention is prepared, it is purged with an inert gas such as nitrogen or argon to maintain a hypoxic condition, and sterilized and filtered.

In addition, the stabilized femetrexed formulations of the present invention may be packaged in a suitable container known in the art. For example, suitable containers may be glass vials, glass bottles, cartridges, pre-filled injections and the like, preferably glass vials.

The present invention Pemetrexed injectable liquid preparation is dispensed in a pre-cleaned sterile container and the surface of the cap suitable for the container is sealed with a Teflon stopper inert to the liquid formulation containing Pemetrexed juice. At this time, if necessary, a space between the injectable liquid preparation and the plug is filled with an inert gas. After attaching the stopper using a crimper, the vial filled with injectable liquid formulation is sterilized by heating if necessary.

The present invention also relates to a pemetrexed preparation comprising, as an active ingredient, pemetrexed or a pharmaceutically acceptable salt thereof, contained in a sealed container, wherein the amount of oxygen in the head space in the container is controlled to 3 v / v% or less To provide a pemetrexed preparation.

As used herein, the term "headspace" refers to the upper space in which the pemetrexed formulation remains filled in a sealed container. The volume of the headspace may vary depending on the total internal volume of the container used and the amount of femetrexed charged.

As used herein, the term "oxygen content in head space" refers to the volume percentage (%) occupied by oxygen in the volume of the total head space. Therefore, the unit of% representing the unit of the oxygen amount means the volume (v / v)%.

In the present invention, the amount of oxygen in the headspace may be 3 v / v% or less, i.e. 0 v / v% to 3 v / v%, specifically 0.001 v / v% to 3 v / v%. In one embodiment, the amount of oxygen in the headspace may be 3 v / v%.

The containers usable in the present invention are as described above, preferably glass vials.

In the present invention, it is preferable that the preparation further comprises N-acetyl-L-cysteine and sodium citrate. At this time, the preferable concentration ratio of the above-mentioned Pemetrexed or its pharmaceutically acceptable salt, N-acetyl-L-cysteine and sodium citrate is as described above.

In the present invention, the preparation is preferably a liquid preparation which can be stored in a solution state, in particular a liquid formulation for injection in a sealed container ready-to-use.

In the present invention, the control of the amount of oxygen in the head space in the vessel can be performed by replacing oxygen in the head space with an inert gas. The inert gas may be nitrogen or argon, but is not limited thereto.

The present invention solves the problem that a pemetrexed preparation containing pemetrexed or a pharmaceutically acceptable salt thereof as an active ingredient contained in a sealed container produces a flexible substance due to oxygen in the head space, The stability of the formulation can be further improved by controlling the oxygen amount in the headspace to 3 v / v% or less (Experimental Example 1, Table 10).

According to the present invention, as described above, a method of controlling the amount of oxygen in the head space, for example, a method of replacing oxygen with nitrogen, can prevent oxidation that may occur in the head space. According to the present invention, an inexpensive and stabilized injectable preparation containing femetrexed can be scaled up to a commercial level and mass-produced using a currently widely used automation system.

The present invention can provide a pemetrexed preparation that is easy to manufacture commercially, can prevent microbial contamination during freeze-drying or reconstitution, and is improved in dosage convenience and stability. In addition, by containing acetylcysteine as an antioxidant and sodium citrate as a buffer, stabilized pemetrexed preparations which are free from abnormalities such as discoloration or precipitate generation compared with conventional liquid preparations containing pemetrexed, . Further, the present invention can provide a more stable pemetrexed formulation by preventing oxidation that can occur in the headspace by controlling the amount of oxygen in the head space.

1 is a graph showing the results of the severe four-week stability of the conventional liquid formulation containing pemetrexed (Comparative Example 1, Comparative Example 12, and Comparative Example 14) and the liquid formulation containing pemetrex according to the present invention (Example 11) (60 ° C / 80%). At this time, the characteristics of the injection water are also shown for comparison.

Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These embodiments are only for illustrating the present invention, and the scope of the present invention is not limited by these embodiments.

Example  1 to 14: acetyl cysteine as an antioxidant, As a buffer  Sodium citrate in different contents Femetrexed  Preparation of solution containing injectable solution

After dissolving 2.5 g of D-mannitol in 100 ml of water for injection, acetylcysteine and sodium citrate were added in order to completely dissolve in the concentration shown in Table 1 below. 2.5 g of pemetrexed was gradually added thereto (Example 14 was gradually added with 3.0 g of pemetrexed), stirred until the solution became transparent, and then adjusted to pH of the following Table 1 using hydrochloric acid or sodium hydroxide aqueous solution I did it. The solution was aseptically filtered through a 0.22 μm membrane filter sterilized in a clean room. The resulting solution was purged with nitrogen in a clean, sterilized, sealable container and filled.

The components and pH of the resulting injectable solution containing pemetrexed were as shown in Table 1 below.

Active ingredient
density
(mg / mL) Antioxidant Antioxidant
density
(mg / mL) Buffer Buffer concentration
(mg / mL) pH Example 1 25 Acetylcysteine 0.15 Sodium citrate 2.0 7.3 Example 2 25 Acetylcysteine 0.3 Sodium citrate 2.0 7.2 Example 3 25 Acetylcysteine 0.5 Sodium citrate 1.0 7.2 Example 4 25 Acetylcysteine 0.5 Sodium citrate 2.0 7.2 Example 5 25 Acetylcysteine 0.5 Sodium citrate 3.0 7.3 Example 6 25 Acetylcysteine 0.5 Sodium citrate 4.0 7.4 Example 7 25 Acetylcysteine 0.5 Sodium citrate 5.0 7.4 Example 8 25 Acetylcysteine 1.0 Sodium citrate 2.0 7.4 Example 9 25 Acetylcysteine 1.5 Sodium citrate 1.0 7.5 Example 10 25 Acetylcysteine 1.5 Sodium citrate 2.0 7.5 Example 11 25 Acetylcysteine 1.5 Sodium citrate 5.0 7.5 Example 12 25 Acetylcysteine 1.5 Sodium citrate 15.0 7.5 Example 13 25 Acetylcysteine 2.0 Sodium citrate 5.0 7.4 Example 14 30 Acetylcysteine 1.5 Sodium citrate 5.0 7.4

Example  15 to 16: pH  According to the present invention, Femetrexed  Manufacture of injectable injection solutions

After dissolving 2.5 g of D-mannitol in 100 ml of water for injection, acetylcysteine and sodium citrate were added in the order of Table 2 to completely dissolve. 2.5 g of pemetrexed was slowly added thereto, stirred until the solution became transparent, and then adjusted to the pH shown in Table 2 using hydrochloric acid or sodium hydroxide aqueous solution. The solution was aseptically filtered through a 0.22 μm membrane filter sterilized in a clean room. The resulting solution was purged with nitrogen in a clean, sterilized, sealable container and filled.

The components and pH of the resulting injectable solution containing pemetrexed were as shown in Table 2 below.

Active ingredient
density
(mg / mL) Antioxidant Antioxidant
Concentration (M) Buffer Buffer concentration
(mg / mL) pH Example 15 25 Acetylcysteine 1.5 Sodium citrate 5.0 6.8 Example 16 25 Acetylcysteine 1.5 Sodium citrate 5.0 7.2

Example  17-20: Headspace  Depending on the oxygen content Femetrexed  Manufacture of injectable injection solutions

After dissolving 2.5 g of D-mannitol in 100 ml of water for injection, acetylcysteine and sodium citrate were added in order to completely dissolve the solution at the concentration shown in Table 3 below. 2.5 g of pemetrexed was slowly added thereto, stirred until the solution became transparent, and adjusted to the pH shown in Table 3 using hydrochloric acid or aqueous sodium hydroxide solution.

The solution was aseptically filtered through a 0.22 μm membrane filter sterilized in a clean room. The resulting solution was purged and purged with nitrogen into the headspace oxygen amount of Table 3 in a clean, sterilized, sealable container.

The components of the resulting injectable solution containing pemetrexed and the headspace oxygen amount were as shown in Table 3 below.

Active ingredient
density
(mg / mL) Antioxidant Antioxidant
Concentration (M) Buffer Buffer concentration
(mg / mL) pH Headspace oxygen content
(v / v%) Example 17 25 Acetylcysteine 1.5 Sodium citrate 5.0 7.4 3 Example 18 25 Acetylcysteine 1.5 Sodium citrate 5.0 7.4 5 Example 19 25 Acetylcysteine - Sodium citrate - 7.4 3 Example 20 25 Acetylcysteine - Sodium citrate - 7.4 5

Comparative Example  1 ~ 8: Depending on the type of antioxidant Femetrexed  Preparation of injection solutions

A solution containing pemetrexed was prepared in the same manner as in Example 1, with the composition and content shown in Table 4 below. In Comparative Example 1, a solution containing only the injected water as a carrier without adding an antioxidant was prepared.

Active ingredient concentration
(mg / mL) Antioxidant type Antioxidant concentration
(mg / mL) pH Comparative Example 1 25 - - 7.5 Comparative Example 2 25 Ascorbic acid 0.3 6.5 Comparative Example 3 25 Sodium thiosulfate 0.3 7.5 Comparative Example 4 25 Butylated hydroxyanisole 0.3 9.9 Comparative Example 5 25 Propyl gallate 0.3 9.5 Comparative Example 6 25 EDTA 0.3 6.8 Comparative Example 7 25 L-cystine 0.3 7.5 Comparative Example 8 25 L-methionine 0.3 7.4

Comparative Example  9 ~ 14: antioxidant type and Buffer  Depending on addition Femetrexed  Preparation of injection solutions

A solution containing pemetrexed was prepared in the same manner as in Example 1, with the composition and content shown in Table 5 below.

Active ingredient
density
(mg / mL) Antioxidant
Kinds Antioxidant
density
(mg / mL) Buffer Buffer concentration
(mg / mL) pH Comparative Example 9 25 - - Sodium citrate 1.5 7.4 Comparative Example 10 25 - - Sodium citrate 2.0 7.3 Comparative Example 11 25 Acetylcysteine 0.5 - - 7.2 Comparative Example 12 25 Acetylcysteine 1.5 - - 7.5 Comparative Example 13 25 L-cysteine 0.5 - - 7.5 Comparative Example 14 25 L-cysteine 1.5 - - 7.5

Comparative Example  15-17: Buffer  Depending on the type Femetrexed  Preparation of injection solutions

A solution containing pemetrexed was prepared in the same manner as in Example 1, with the composition and content shown in Table 6 below.

Active ingredient
density
(mg / mL) Antioxidant
Kinds Antioxidant
density
(mg / mL) Buffer Buffer concentration
(mg / mL) pH Comparative Example 15 25 Acetylcysteine 1.5 Sodium acetate 5.0 7.4 Comparative Example 16 25 Acetylcysteine 1.5 Sodium hydrogen phosphate 5.0 7.5 Comparative Example 17 25 Acetylcysteine 1.5 Potassium dihydrogen phosphate 5.0 7.5

Experimental Example : Stability test

The compositions prepared in Examples 1 to 20 and Comparative Examples 1 to 17 were subjected to a stability test under a severe condition (60 ° C / 80%) for 4 weeks. Of these, Example 11 and Comparative Examples 1, 12 and 14 were subjected to a stability test for four months under accelerated conditions (40 ° C / 70%). The stability evaluation was carried out by measuring the content of the residual amount of pemetrexed and the content of the flexible substance in the aqueous solution under the conditions described in Table 7 using the high performance liquid chromatography method.

column C8, 150 x 4.6 mm, 3.5 m Detector Ultraviolet absorption spectrophotometer (250 nm) Mobile phase Gradient method
Mobile Phase A-Acetic Acid Buffer: Acetonitrile (97: 3)
Mobile Phase B-Acetic Acid Buffer: Acetonitrile (87.5: 12.5)
Acetic acid buffer solution (0.03 mol / L, pH 5.5) 3.4 mL of acetic acid (100%) was added to 2 L of water, and the pH was adjusted to 5.5 with 50% sodium hydroxide after stirring Flow rate 1 mL / min Column temperature 35 ℃ Analysis time 55 minutes

Experimental Example 1: Harshness stability test (60 占 폚 / 80%, 4 weeks stability evaluation)

The results of conducting the severe stability test as described above are shown in Tables 8 to 13 below.

In addition, the formulation of Example 11 and the conventional liquid formulation containing pemetrexed as Comparative Examples 1, 2, and 3 were used as the injectable liquid preparation containing pemetrex according to the present invention among the liquid preparations containing pemetrexed prepared above, (60 ° C / 80%) of the preparation of Comparative Example 12 and Comparative Example 14 is shown in FIG. At this time, the characteristics of the injection water are also shown for comparison.

Active ingredient
density
(mg / mL) Antioxidant Antioxidant
density
(mg / mL) Buffer Buffer
density
(mg / mL) pH Flexible Substance (%)
Individual / total Example 1 25 Acetylcysteine 0.15 Sodium citrate 2.0
7.3 Early 0.05 / 0.1 (colorless transparent) 2 weeks 0.09 / 0.3 (yellow) 4 weeks 0.11 / 0.6 (yellow) Example 2 25 Acetylcysteine 0.3 Sodium citrate 2.0 7.2 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.10 / 0.3 (light yellow) 4 weeks 0.12 / 0.6 (yellow) Example 3 25 Acetylcysteine 0.5 Sodium citrate 1.0 7.2 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.10 / 0.1 (light yellow) 4 weeks 0.11 / 0.4 (light yellow) Example 4 25 Acetylcysteine 0.5 Sodium citrate 2.0 7.2 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.10 / 0.1 (light yellow) 4 weeks 0.13 / 0.5 (light yellow) Example 5 25 Acetylcysteine 0.5 Sodium citrate 3.0 7.3 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.15 / 0.3 (light yellow) 4 weeks 0.10 / 0.4 (light yellow) Example 6 25 Acetylcysteine 0.5 Sodium citrate 4.0 7.4 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.10 / 0.2 (colorless transparent) 4 weeks 0.12 / 0.5 (light yellow) Example 7 25 Acetylcysteine 0.5 Sodium citrate 5.0 7.4 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.10 / 0.2 (colorless transparent) 4 weeks 0.12 / 0.5 (light yellow) Example 8 25 Acetylcysteine 1.0 Sodium citrate 2.0 7.4 Early 0.08 / 0.1 (colorless transparent) 2 weeks 0.11 / 0.2 (colorless transparent) 4 weeks 0.13 / 0.5 (colorless transparent) Example 9 25 Acetylcysteine 1.5 Sodium citrate 1.0 7.5 Early 0.08 / 0.1 (colorless transparent) 2 weeks 0.11 / 0.2 (colorless transparent) 4 weeks 0.10 / 0.3 (colorless transparent) Example 10 25 Acetylcysteine 1.5 Sodium citrate 2.0 7.5 Early 0.08 / 0.1 (colorless transparent) 2 weeks 0.12 / 0.1 (colorless transparent) 4 weeks 0.12 / 0.3 (colorless transparent) Example 11 25 Acetylcysteine 1.5 Sodium citrate 5.0 7.5 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.10 / 0.1 (colorless transparent) 4 weeks 0.11 / 0.2 (colorless transparent) Example 12 25 Acetylcysteine 1.5 Sodium citrate 15.0 7.5 Early 0.07 / 0.1 (colorless transparent) 2 weeks 0.11 / 0.1 (colorless transparent) 4 weeks 0.11 / 0.2 (colorless transparent) Example 13 25 Acetylcysteine 2.0 Sodium citrate 5.0 7.4 Early 0.20 / 0.2 (colorless transparent) 2 weeks 0.25 / 0.4 (light yellowish yellow) 4 weeks 0.25 / 0.6 (light yellow) Example 14 30 Acetylcysteine 1.5 Sodium citrate 5.0 7.5 Early 0.11 / 0.1 (colorless transparent) 2 weeks 0.15 / 0.2 (colorless transparent) 4 weeks 0.15 / 0.3 (light yellowish yellow)

As can be seen from the results in Table 8, when 1.5 mg / mL of acetylcysteine as an antioxidant and 1 to 15 mg / mL of sodium citrate as a buffer were included together, Hereinafter, the total flexible material showed excellent stability at 1.0% or less (Examples 9 to 12).

When the concentration of sodium citrate was 2.0 mg / mL when the concentration of acetylcysteine was 1.0 to 1.5 mg / mL, the stability of the individual soft materials was 0.2% or less and the total softness was 1.0% or less (Example 8 and 10). When the concentration of acetylcysteine was lower than this, it was confirmed that the flexible material met the standard, but the change in properties (discoloration) occurred (Examples 1 and 2).

In addition, when the concentration of acetylcysteine was 2.0 mg / mL and the concentration of sodium citrate was 5.0 mg / mL, it was confirmed that the supernatant was in conformity with the standard, but the change in properties (discoloration) occurred (Example 13).

In addition, when the concentration of acetylcysteine was 0.5 mg / mL and the concentration of sodium citrate was 1.0 to 5.0 mg / mL, it was confirmed that the substance conformed to the standard, but a change in properties (discoloration) occurred (Example 3 7).

Active ingredient
density
(mg / mL) Antioxidant Antioxidant
density
(mg / mL) Buffer Buffer
density
(mg / mL) pH Flexible Substance (%)
Individual / total Example 15 25 Acetyl
Cysteine 1.5 Sodium citrate 5.0 6.9 Early 0.16 / 0.2
(transparent) 2 weeks 0.18 / 0.2
(transparent) 4 weeks 0.17 / 0.3
(transparent) Example 16 25 Acetyl
Cysteine 1.5 Sodium citrate 5.0 7.2 Early 0.14 / 0.2
(transparent) 2 weeks 0.15 / 0.3
(transparent) 4 weeks 0.14 / 0.2
(transparent) Example 11 25 Acetyl
Cysteine 1.5 Sodium citrate 5.0 7.5 Early 0.07 / 0.1
(transparent) 2 weeks 0.10 / 0.1
(transparent) 4 weeks 0.11 / 0.2
(transparent)

As can be seen from the results in Table 9, when acetylcysteine was included as an antioxidant and sodium citrate as a buffer, the individual flexible substances were 0.2% or less and the total amount of the flexible substances 1.0% or less.

available
ingredient
density
(mg / mL) Antioxidant Antioxidant
density
(mg / mL) Buffer Buffer
density
(mg / mL) pH head
space
Oxygen amount
(v / v%) Flexible Substance (%)
Individual / total Example 17 25 Acetyl
Cysteine 1.5 Sodium citrate 5.0 7.4 3 Early 0.05 or less /0.0
(transparent) 2 weeks 0.05 or less /0.0
(transparent) 4 weeks 0.05 or less /0.0
(transparent) Example 18 25 Acetyl
Cysteine 1.5 Sodium citrate 5.0 7.4 5 Early 0.05 or less /0.0
(transparent) 2 weeks 0.12 / 0.4
(transparent) 4 weeks 0.22 / 0.7
(transparent) Example 19 25 Acetyl
Cysteine - Sodium citrate - 7.4 3 Early
0.05 or less /0.0
(transparent) 2 weeks
0.18 / 0.7
(Light yellow) 4 weeks 0.57 / 1.5
(yellow) Example 20 25 Acetyl
Cysteine - Sodium citrate - 7.4 5 Early
0.05 or less /0.0
(transparent) 2 weeks
0.25 / 0.8
(Light yellow) 4 weeks 0.61 / 1.8
(yellow)

When the preparation is mass-produced for commercialization, the opportunity for oxygen contact, which is a major source of the flexible substance, may increase. In order to prevent this, the stability of the formulation can be ensured by controlling the residual amount of oxygen in the process.

As can be seen from the results in Table 10, when the head space oxygen amount was controlled to be lower than the conventional manufacturing level (Example 11) when the acetyl cysteine as an antioxidant and sodium citrate as a buffer were included together, Indicating that it is superior in terms of stability (Examples 17 to 18). Particularly, when the oxygen content in the head space was controlled to 3 v / v% or less, the stability of the individual flexible material was 0.2% or less and the total flexible material was 1.0% or less during the four weeks of stability.

Even when the antioxidant was not added, it was confirmed that the stability was secured for up to two weeks by controlling the oxygen amount in the head space to 3 v / v% or less, as compared with the case where the oxygen amount in the head space was not controlled (Comparative Example 1) Example 19). This indicates that control of the amount of oxygen in the headspace contributes to improvement of formulation stability, but more complete stability was obtained in the case of addition of an antioxidant and a buffer (Examples 17 and 18).

Active ingredient
density
(mg / mL) Antioxidant
Kinds Antioxidant
density
(mg / mL) pH Flexible Substance (%)
[Individual / total] Comparative Example 1 25 - - 7.5 Early 0.01 / 0.1 2 weeks 0.33 / 0.8 4 weeks 0.66 / 2.1 Comparative Example 2 25 Ascorbic acid 0.3 6.5 Early 0.63 / 0.7 2 weeks 0.6 / 1.0 4 weeks Not measurable
(Unstable) Comparative Example 3 25 Sodium thiosulfate 0.3 7.5 Early 0.04 / 0.2 2 weeks 0.24 / 0.6 4 weeks 0.72 / 1.7 Comparative Example 4 25 Butylated
Hydroxy
Anisol 0.3 9.9 Early 0.1 / 0.3 2 weeks 0.45 / 1.3 4 weeks Not measurable
(Unstable) Comparative Example 5 25 Propyl gallate 0.3 9.5 Early 0.11 / 0.4 2 weeks 1.03 / 2.7 4 weeks Not measurable
(Unstable) Comparative Example 6 25 EDTA 0.3 6.5 Early 0.53 / 2.7 2 weeks 1.23 / 3.0 4 weeks Not measurable
(Unstable) Comparative Example 7 25 L-cystine 0.3 7.5 Early 0.05 / 0.2 2 weeks 0.38 / 1.0 4 weeks 0.71 / 1.8 Comparative Example 8 25 L-methionine 0.3 7.4 Early 0.05 / 0.2 2 weeks 0.35 / 0.9 4 weeks 0.51 / 1.3

As can be seen from the results of Table 11, when the conventional antioxidants are included, the change in properties such as precipitation or discoloration during the period of severe stability, the increase in the content of the individual softeners by 0.2% or more, the increase of the total softenes by more than 1.0% . Thus, when injection solutions containing pemetrexed were prepared using a commonly used antioxidant, it was found that the solution exhibited unsuitable stability in terms of changes in the shape of the suppositories and changes in properties.

Active ingredient
density
(mg / mL) Antioxidant
Kinds Antioxidant
density
(mg / mL) Buffer Buffer
density
(mg / mL) pH Flexible Substance (%)
Individual / total Comparative Example 9 25 -      - Sodium citrate 1.5 7.4 Early 0.15 / 0.2
(transparent) 7.4 2 weeks 0.13 / 0.6
(Dark yellow) - 4 weeks Not measurable
(Unstable) Comparative Example 10 25 - - Sodium citrate 2.0 7.3 Early 0.03 / 0.0
(transparent) 7.0 2 weeks 0.11 / 0.3
(Dark yellow) 7.0 4 weeks 0.17 / 0.5
(Dark yellow) Example 11 25 Acetyl
Cysteine 1.5 Sodium citrate 5.0 7.5 Early 0.07 / 0.1
(transparent) 7.3 2 weeks 0.10 / 0.1
(transparent) 7.2 4 weeks 0.11 / 0.2
(transparent) Comparative Example 11 25 Acetyl
Cysteine 0.5 - - 7.2 Early 0.12 / 0.1
(transparent) 6.9 2 weeks 0.14 / 0.3
(Light yellow) 6.9 4 weeks 0.15 / 0.5
(Light yellow) Example 8 25 Acetyl
Cysteine 0.5 Sodium citrate 5.0 7.4 Early 0.07 / 0.1
(transparent) 7.1 2 weeks 0.10 / 0.2
(transparent) 7.1 4 weeks 0.12 / 0.5
(Light yellow) Comparative Example 12 25 Acetyl
Cysteine 1.5 - - 7.5 Early 0.09 / 0.1
(transparent) 7.1 2 weeks 0.12 / 0.3
(transparent) 7.0 4 weeks 0.12 / 0.3
(Light yellow) Comparative Example 13 25 L-cysteine 0.5 - - 7.5 Early 0.12 / 0.2
(transparent) 6.9 2 weeks 0.12 / 0.2
(Light yellow and odor is generated) 6.9 4 weeks 0.12 / 0.3
(Light yellow) Comparative Example 14 25 L-cysteine 1.5 - - 7.5 Early 0.09 / 0.1
(transparent) 7.4 2 weeks 0.11 / 0.1
(A sense of smell is generated) 7.2 4 weeks 0.12 / 0.2
(A sense of smell is generated)

As can be seen from the results of Table 12, when acetyl cysteine as an antioxidant and sodium citrate as a buffer are included together, it is preferable that the concentration of acetylcysteine is 1.5 mg / mL and the concentration of sodium citrate is 5.0 mg / mL , It was found that the stability of the individual flexible substance was 0.2% or less and the total amount of the flexible substance was 1.0% or less during the 4-week stability period (Example 11).

On the other hand, when only sodium citrate as a buffer was included, it was observed that the color of the solution changed from deeply dark to dark yellow (Comparative Examples 9 and 10). In addition, even when the antioxidants L-cysteine and acetylcysteine alone were included, the color of the solution turned to be yellowish from the 2nd week of the harsh period, and it was observed that the pH dropped remarkably during the harsh 4-week period Example 11, and Comparative Examples 12 to 14). That is, unlike the case of using only acetylcysteine or sodium citrate alone, the stability of the substance was remarkably improved by using both of them.

In addition, in Comparative Examples 13 and 14 including only L-cysteine, a feeling of deodorant similar to that of egg rot was generated from the 2nd week of the severe case.

Active ingredient
density
(mg / mL) Antioxidant
Kinds Antioxidant
density
(mg / mL) Buffer Buffer
density
(mg / mL) pH Flexible Substance (%)
Individual / total Example 11 25 Acetyl
Cysteine 1.5 Sodium citrate 5.0 7.5 Early 0.07 / 0.1
(transparent) 7.3 2 weeks 0.10 / 0.1
(transparent) Comparative Example 15 25 Acetyl
Cysteine 1.5 Sodium acetate 5.0 7.4 Early 0.89 / 1.45
(Light yellow) 7.1 2 weeks 0.86 / 2.3
(Dark yellow) Comparative Example 16 25 Acetyl
Cysteine 1.5 Sodium hydrogen phosphate 5.0 7.5 Early 0.23 / 0.2
(transparent) 7.4 2 weeks 0.21 / 0.9
(yellow) Comparative Example 17 25 Acetyl
Cysteine 1.5 Potassium dihydrogen phosphate 5.0 7.5 Early 0.24 / 0.2
(transparent) 7.2 2 weeks 0.23 / 0.8
(Light yellow)

As can be seen from the results of Table 13, when comparing the stability results of various antioxidants including acetylcysteine and various buffers generally used as buffers, when the remaining buffer except for sodium citrate is used, It was observed that the change (discoloration), the content of individual softeners increased by more than 0.2%, and the content of total softeners increased by more than 1.0%.

Therefore, it was found that when the injection solution containing pemetrexed was prepared, the best stability was obtained only when acetyl cysteine was used as an antioxidant and sodium citrate was used as a buffer.

Experimental Example  2: Accelerated stability test (40 占 폚 / 70%, 4 month stability evaluation)

The results of the acceleration stability test as described above are shown in Table 14 below.

Active ingredient
density
(mg / mL) Antioxidant
Kinds Antioxidant
density
(mg / mL) Buffer Buffer
density
(mg / mL) pH Flexible Substance (%)
Individual / total Example 11 25 Acetylcysteine 1.5 Sodium citrate 5.0 7.5 Early 0.07 / 0.1
(transparent) 7.3 2 months 0.10 / 0.1
(transparent) 7.3 4 months 0.14 / 0.2
(transparent) 7.2 6 months 0.18 / 0.4
(Light yellow) Comparative Example 1 25 - - - - 7.5 Early 0.01 / 0.1 - 2 months 0.04 / 0.1
(yellow) - 4 months 0.60 / 1.5
(yellow) Comparative Example 12 25 Acetylcysteine 1.5 - - 7.5 Early 0.09 / 0.1
(transparent) 7.1 4 months 0.33 / 0.8
(transparent) Comparative Example 14 25 L-cysteine 1.5 - - 7.5 Early 0.09 / 0.1
(transparent) 7.2 4 months 0.12 / 0.2
(Colorless transparent and odor sensation occurs)

As can be seen from the results in Table 14, in the case of Comparative Example 1 prepared by using only the injected water as the carrier without the addition of the antioxidant, the property of the solution changed to yellow from 2 months after the acceleration, 0.2% or more, and 1% or more of total flexible materials. However, in Example 11 including acetylcysteine as an antioxidant and sodium citrate as a buffer, the stability of the individual flexible substance was 0.2% or less and the total amount of the flexible substance was 1.0% or less during the accelerated 6-month stability period.

On the other hand, even when the antioxidant, acetylcysteine alone was included, the color of the solution turned yellow from 4 months after the acceleration, and the individual soft materials were found to be not more than 0.2% (Comparative Example 12).

In addition, in the case of Comparative Example 14 containing only L-cysteine, the flexible material was in conformity with the standard, but as in the case of the harsh stability result, a sense of deodorant similar to the rotten smell of the egg occurred from four months after the acceleration. However, , There was no such feeling.

Claims (13)

A femetrexed preparation comprising, as an active ingredient, Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate.
2. The formulation of claim 1, wherein the concentration ratio of Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate is 1 to 30: 0.15 to 2.0: 1.0 to 15.0.
The formulation according to claim 2, wherein the concentration ratio of Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate is 1 to 30: 1.5: 1.0 to 15.0.
The formulation according to claim 1, wherein the preparation is a liquid preparation that can be stored in a solution state.
The formulation of claim 1, wherein the formulation is a injectable liquid formulation in a ready-to-use sealed container.
Claims 1. A Pemetrexed formulation comprising Pemetrexed or a pharmaceutically acceptable salt thereof as an active ingredient in a sealed container, wherein the formulation is such that the amount of oxygen in the head space in the container is at least 3 v / v ≪ / RTI > or less.
7. The formulation of claim 6, wherein the formulation further comprises N-acetyl-L-cysteine and sodium citrate.
8. The formulation according to claim 7, wherein the concentration ratio of Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate is 1 to 30: 0.15 to 2.0: 1.0 to 15.0.
9. The formulation according to claim 8, wherein the concentration ratio of Pemetrexed or a pharmaceutically acceptable salt thereof, N-acetyl-L-cysteine and sodium citrate is 1 to 30: 1.5: 1.0 to 15.0.
The formulation according to claim 6, wherein the preparation is a liquid preparation that can be stored in a solution state.
7. The formulation of claim 6, wherein the formulation is a injectable liquid formulation in a ready-to-use sealed container.
7. The formulation according to claim 6, wherein the formulation is prepared by replacing oxygen in the head space in the vessel with an inert gas to adjust the amount of oxygen.
13. The formulation of claim 12, wherein the inert gas is nitrogen or argon.

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